Difference between revisions of "Part:BBa K1033206"
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<partinfo>BBa_K1033206 short</partinfo> | <partinfo>BBa_K1033206 short</partinfo> | ||
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The backbone pSBLbC is a shuttle vector between E. coli, Lacotbacillus, and some other lactic acid bacteria. It has been used for subcloning in E. coli and to express a chromoprotein [[Part:BBa_K1033209|amilCP]]. For use in Lactobacillus we recommend the version with erythromycin resistance ([[Part:BBa_K1033207|pSBLbE]]) since that has been successfully used to transform Lactobacillus. | The backbone pSBLbC is a shuttle vector between E. coli, Lacotbacillus, and some other lactic acid bacteria. It has been used for subcloning in E. coli and to express a chromoprotein [[Part:BBa_K1033209|amilCP]]. For use in Lactobacillus we recommend the version with erythromycin resistance ([[Part:BBa_K1033207|pSBLbE]]) since that has been successfully used to transform Lactobacillus. | ||
https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png | https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png | ||
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<i><b>Fig 1:</b> E. coli D5-alpha with pSBLbC to which amilCP and lactobacillus promotor CP29 have been subcloned</i> | <i><b>Fig 1:</b> E. coli D5-alpha with pSBLbC to which amilCP and lactobacillus promotor CP29 have been subcloned</i> | ||
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We also had to change the promotor of the chloramphenicol resistance cassette to one that would initiate transcription effectively in <i>Lactobacillus</i> we tried constitutive promotors but finally got [[Part:BBa_K1033222|CP29]] to work. | We also had to change the promotor of the chloramphenicol resistance cassette to one that would initiate transcription effectively in <i>Lactobacillus</i> we tried constitutive promotors but finally got [[Part:BBa_K1033222|CP29]] to work. | ||
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Revision as of 22:05, 3 October 2013
Lactobacillus shuttle vector pSBLbC
The backbone pSBLbC is a shuttle vector between E. coli, Lacotbacillus, and some other lactic acid bacteria. It has been used for subcloning in E. coli and to express a chromoprotein amilCP. For use in Lactobacillus we recommend the version with erythromycin resistance (pSBLbE) since that has been successfully used to transform Lactobacillus.
Fig 1: E. coli D5-alpha with pSBLbC to which amilCP and lactobacillus promotor CP29 have been subcloned
It was made by replacing the replicon of BioBrick compatible plasmid pSB4C15 (BBa_K864001) with a broad range replicon from the engineered plasmid pJP059. The replicon is also referred to as pSH71 and is related to pWV01 and that family of rolling circle replicating plasmids.
We also had to change the promotor of the chloramphenicol resistance cassette to one that would initiate transcription effectively in Lactobacillus we tried constitutive promotors but finally got CP29 to work.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 3036
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3036
Illegal NheI site found at 1981
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 3042 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3036
Illegal BamHI site found at 1960 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 3036
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 3036
Illegal XbaI site found at 3051
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000COMPATIBLE WITH RFC[1000]